1. What is different about the Latex Conjugation Kit?
The latex particles are specially treated to enhance the handling of the latex and permit easy covalent attachment of antibodies and other proteins. The conjugation reaction is initiated simply by adding a solution of the antibody to the freeze-dried powder; the hands on time is 3 minutes and the conjugate is ready to use within 35 minutes.
Latex nanoparticles are prone to aggregation both before and after conjugation, and harsh methods such as sonication or vortexing are routinely needed to resuspend both conjugated and unconjugated latex. Our special treatment significantly improves the handling of the latex and vastly reduces the propensity of the latex to aggregate. Harsh resuspension methods are not required, and recovery of usual mono-disperse latex particles is very high.
2. What type of linkage to the latex is formed?
The antibody becomes covalently attached to the specially treated surface via lysine residues.
3. Does the conjugated antibody bind to the latex surface?
The special treatment shields the latex surface, preventing surface – antibody interactions. For this reason you cannot use our latex for passive conjugation.
4. How many latex beads are in each kit?
Each vial of freeze dried latex will provide 40ul 1% conjugated latex. In the 4 reaction kit there is 160ul 1% latex in total and in the 10 reaction kit there is a total of 400ul 1% latex. 40ul 1% latex should provide enough conjugate to perform approximately 50 lateral flow assays.
5. What if I need bulk material?
The Mini kits are a convenient for rapid production of small quantities of conjugate for screening purposes. The Midi kits are ten times the size of the Mini kits for medium scale work (e.g. ~500 lateral flow tests). Bulk quantities can also be produced to meet your custom needs. Please contact us to discuss your bulk or custom conjugation needs or if you require any additional information.
6. Can antibodies from different species be used?
Yes. We have tested a variety of species including mouse & goat.
7. Can monoclonal and polyclonal antibodies be used?
Yes. Our Latex Conjugation Kit has been developed for use with both monoclonal and polyclonal antibodies.
8. Can antibody fragments and non-antibody proteins be conjugated?
One of the advantages of the protective coat is that it is less likely than a bare surface to cause denaturation of antibody fragments and other proteins.
9. Is the kit suitable for conjugating small molecules including analytes?
The Latex Conjugation Kit has been optimised for use with antibodies, but it may be possible to conjugate small molecules with primary amine functional groups. Please contact our Technical Support at firstname.lastname@example.org for information and support.
10. What can I do if my antibody is not in a compatible buffer?
The conjugation reaction is sensitive to a number of common buffers and buffer additives. The antibody should be in 10 – 50 mM MES, MOPS or HEPES at pH 6 – 7 for conjugation. As these are not common storage buffers for antibodies and other proteins we have developed the companion product the Antibody Concentration & Clean Up Kit for Latex to switch the antibody buffer to one that is optimal for conjugation efficiency.
11. What can I do if my antibody has non-antibody protein components or is not purified?
To remove contaminating proteins before conjugation or to purify your antibody from ascites fluid, serum or tissue culture supernatant please see our AbPure range of purification products.
12. What colour latex should I buy?
Our blue, red and black latex all behave in a very similar way chemically, so the ideal latex colour will depend on your application and the preference of the scientist.
13. How can I know if my conjugation was a success?
Confirm the success of your antibody conjugation with our Check&Go! kit, a nitrocellulose membrane containing a ‘Test line’ of immobilized Protein A and Protein G called a “half strip”. The Conjugate Check&Go! strips allow visualization of conjugates generated using our beads and nanoparticles at different concentrations of antibody.
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